Pharmaceutical composition comprising botulinum, a non ionic surfactant, sodium chloride and sucrose

ABSTRACT

The invention relates to a solid or liquid pharmaceutical composition comprising botulinum neurotoxin complex (type A, B, C, D, E, F or G) or high purity botulinum neurotoxin (type A, B, C, D, E, F or G), and a surfactant. In particular the invention relates to a solid or liquid pharmaceutical composition comprising a crystalline agent.

The invention relates to a pharmaceutical composition containingbotulinum neurotoxin.

The presently most used botulinum neurotoxin is botulinum neurotoxintype A. This neurotoxin is produced during fermentation in the presenceof Clostridium botulinum strains. Botulinum neurotoxin type A complexes(which include botulinum neurotoxin type A and at least anothernon-toxic protein) are active principles widely used in modern medicine.An example of a pharmaceutical composition based on such a complex isthe product Dysport® currently sold by the company of the Applicants.Among the most common medical indications for which a botulinumneurotoxin type A complex could be used, one could mention the treatmentof a number of muscle disorders (e.g. blepharospasm, hemifacial spasm,torticollis, spasticity, tension headache, back pain or wrinkles), aswell as other disorders such as migraine. Alternatively, high puritybotulinum toxin (i.e. botulinum neurotoxin free from its complexingnon-toxic proteins) may replace the corresponding botulinum toxincomplex as disclosed in PCT applications WO 96/11699 or WO 97/35604.

Currently, the marketed botulinum neurotoxin compositions contain humanserum albumin. However, some concerns have been expressed about albumin(see e.g. in PCT application WO 01/58472). For this reason, thepharmaceutical industry is now considering to find alternativestabilising agents to albumin by other stabilising agents inpharmaceutical compositions.

A possible solution is disclosed in PCT patent application WO 01/58472.In this document, albumin is replaced by a polysaccharide, i.e. apolymer of more than two saccharide molecule monomers, which plays therole of the stabiliser in the botulinum neurotoxin composition.

An alternative solution is the one described in PCT patent applicationWO 97/35604 or U.S. Pat. Nos. 5,512,547 and 5,756,468. In thesedocuments, it is disclosed that pure botulinum neurotoxin (i.e.botulinum neurotoxin free from its complexing non-toxic proteins) can bestabilised by trehalose.

The Applicant has unexpectedly discovered that a surfactant possessessufficient stabilising effects to replace albumin, the polysaccharide ofPCT patent application WO 01/58472 or the trehalose of PCT patentapplication WO 97/35604 in botulinum neurotoxin compositions.

The invention therefore pertains to the use of a surfactant forstabilising a solid or liquid pharmaceutical composition that containsas active principle a botulinum toxin.

By botulinum toxin should be understood a naturally occurring botulinumtoxin or any recombinantly produced botulinum toxin.

By naturally occurring botulinum toxin should be understood either ahigh purity botulinum neurotoxin derived from Clostridium spp or abotulinum neurotoxin complex derived from Clostridium spp.

By high purity botulinum neurotoxin (type A, B, C, D, E, F or G) ismeant, in the present application, botulinum neurotoxin (type A, B, C,D, E, F or G) outside from complexes including at least another protein.In other words, a high purity botulinum neurotoxin (type A, B, C, D, E,F or G) does not contain significant quantities of any other Clostridiumspp derived protein than botulinum neurotoxin (type A, B, C, D, E, F orG).

Preferably, according to the present invention, botulinum neurotoxincomplexes and high purity botulinum neurotoxins will be selected fromthe group consisting of botulinum neurotoxin complex and high puritybotulinum neurotoxin of type A, botulinum neurotoxin complex and highpurity botulinum neurotoxin of type B and botulinum neurotoxin complexand high purity botulinum neurotoxin of type F. More preferably,botulinum neurotoxin complexes and high purity botulinum neurotoxinswill be selected from the group consisting of botulinum neurotoxincomplex and high purity botulinum neurotoxin of type A and botulinumneurotoxin complex and high purity botulinum neurotoxin of type F. Moreparticularly, botulinum neurotoxin complexes and high purity botulinumneurotoxins will be botulinum neurotoxin complexes and high puritybotulinum neurotoxins of type A.

By type A botulinum neurotoxin should be understood any botulinum toxinof type A, and notably botulinum neurotoxins of type A1, A2 or A3. Thesame applies mutatis mutandis to the other serotypes of toxins.

The high purity botulinum neurotoxin (type A, B, C, D, E, F or G) usedaccording to the invention or contained in the above describedpharmaceutical compositions can easily be obtained from thecorresponding botulinum neurotoxin complex, for example as explained inCurrent Topics in Microbiology and Immunology (1995), 195, p. 151-154.High purity Clostridium botulinum toxin (type A, B, C, D, E, F or G) isobtained, for example, by purification of an adequate fermentationmedium (for example, an enriched meat media broth containing ClostridiumBotulinum and left for fermentation—this broth may be, for example, theone described in Current Topics in Microbiology and Immunology (1995),195, p. 150 and DasGupta, “Microbial food toxicants. Clostridiumbotulinum toxins. CRC handbook of foodborne diseases of biologicalorigin”, CRC Boca Raton, p. 25-56). When including high purity botulinumneurotoxin in a composition according to the instant invention, thepurity degree of the toxin should preferably be higher than 80%, morepreferably higher than 90 or 95% and in a more particularly preferredmanner higher than 93% or 99%. It can be assessed, for example, by usingthe purity assay described in the present application.

The instant invention also relates to a solid or liquid pharmaceuticalcomposition comprising:

-   -   (a) a botulinum toxin, and    -   (b) a surfactant.

According to a particular variant of the invention, the pharmaceuticalcomposition will be a solid pharmaceutical composition and willessentially consist of:

-   -   (a) a botulinum toxin, and    -   (b) a surfactant.

According to another particular variant of the invention, thepharmaceutical composition will be a liquid pharmaceutical compositionand will essentially consist of:

-   -   (a) a botulinum toxin,    -   (b) a surfactant, and    -   (c) water.

In the abovementioned pharmaceutical compositions, the surfactant willbe such that it stabilises the botulinum toxin.

A solid pharmaceutical composition according to the invention can beobtained for example by lyophilising a sterile water solution containingthe components (a) and (b) as mentioned previously. A liquidpharmaceutical composition according to the invention will be obtainedby mixing the solid (e.g. lyophilised) mixture of components (a) and (b)with sterile water.

According to the invention, the concentrations of said components (a)and (b) in the solution to be lyophilised or the liquid pharmaceuticalcomposition will preferably be as follows:

the solution will contain from 50 to 10,000 LD50 units of botulinumneurotoxin complex (type A, B, C, D, E, F or G) or high purity botulinumneurotoxin (type A, B, C, D, E, F or G) per ml of solution, preferably[the solution will contain] from 50 to 3,000 LD50 units of botulinumneurotoxin complex (type A, B, C, D, E, F or G) or high purity botulinumneurotoxin (type A, B, C, D, E, F or G) per ml of solution, morepreferably from 100 to 2,500 LD50 units of botulinum neurotoxin complex(type A, B, C, D, E, F or G) or high purity botulinum neurotoxin (typeA, B, C, D, E, F or G) per ml of solution and most preferably from 100to 2,000 LD50 units of botulinum neurotoxin complex (type A, B, C, D, E,F or G) or high purity botulinum neurotoxin (type A, B, C, D, E, F or G)per ml of solution;

the concentration of surfactant will be from above critical micellarconcentration to a concentration of 1% v/v, and notably from about0.005% to 0.02% v/v in the case of polysorbate 80.

Preferably, the surfactant will be a non-ionic surfactant. Non-ionicsurfactants include notably polysorbates and block copolymers likepoloxamers (i.e. copolymers of polyethylene and propylene glycol).According to a preferred variant of the invention, the surfactant willbe a polysorbate. More preferably, a polysorbate included in acomposition according to the instant invention will have a meanpolymerisation degree of from 20 to 100 monomer units (preferably about80), and may for example be polysorbate 80. Preferably also, thepolysorbate should be vegetable-derived.

According to a preferred execution mode of the invention, the solid orliquid pharmaceutical composition will also contain a crystalline agent.

By crystalline agent is meant an agent which, inter alia, would maintaina mechanically strong cake structure to lyophilised botulinum neurotoxincomplex (type A, B, C, D, E, F or G) or high purity botulinum neurotoxin(type A, B, C, D, E, F or G). When included in solid formulations,crystalline agents also have a bulking effect. Crystalline agentsnotably include sodium chloride. Contrarily to what was taught in theprior art (see e.g. Goodnough, M. C. and Johnson, E. A., Applied andEnvironmental Microbiology (1992), 58(10), 3426-3428), the use of sodiumchloride for this type of compositions further improves the stability ofthe botulinum toxin composition.

According to yet another preferred execution mode of the invention, thesolid or liquid pharmaceutical composition will also contain a buffer tomaintain pH from 5.5 to 7.5.

The buffer can be any buffer able to maintain the adequate pH.Preferably, the buffer for compositions according to the invention willbe chosen from the group consisting of succinate and an amino acid likehistidine. In particular, the buffer will be histidine. Preferably, thepH will be at least equal to 5.5 or 5.8, and most preferably at leastequal to 6.0 or 6.5. Preferably also, the pH will be equal to or lessthan 7.5 or 7.0, more preferably equal to or less than 6.8.

Preferably, the solid or liquid pharmaceutical composition of theinvention may also contain a disaccharide.

The disaccharide used in compositions according to the invention willpreferably be chosen from the group consisting of sucrose, trehalose,mannitol and lactose. The disaccharide used in compositions according tothe invention will more preferably be chosen from the group consistingof sucrose and trehalose. In particular, the disaccharide used incompositions according to the invention will be sucrose. Preferably, thedisaccharide will be present in the pharmaceutical compositions of theinstant invention, particularly when the compositions are in a solidform.

The instant invention therefore notably relates to a solid or liquidpharmaceutical composition comprising:

-   -   (a) botulinum neurotoxin complex (type A, B, C, D, E, F or G) or        high purity botulinum neurotoxin (type A, B, C, D, E, F or G),    -   (b) a surfactant,    -   (c) a crystalline agent,    -   (d) a buffer to maintain pH between 5.5 to 7.5.

Preferably, a disaccharide will also be included in the pharmaceuticalcompositions according to the present invention, especially when theyare in a solid form.

According to this variant of the invention, a solid pharmaceuticalcomposition can be obtained by lyophilising a sterile water solutioncontaining the components (a) to (d) as mentioned previously. A liquidpharmaceutical composition according to the invention will be obtainedby mixing a solid (e.g. lyophilized) mixture of said components (a) to(d) with sterile water.

According to the invention, the concentrations of said components (a) to(d) in the solution to be lyophilised or the liquid pharmaceuticalcomposition will preferably be as follows:

the solution will contain from 50 to 10,000 LD50 units of botulinumneurotoxin complex (type A, B, C, D, E, F or G) or high purity botulinumneurotoxin (type A, B, C, D, E, F or G) per ml of solution, preferably[the solution will contain] from 50 to 3,000 LD50 units of botulinumneurotoxin complex (type A, B, C, D, E, F or G) or high purity botulinumneurotoxin (type A, B, C, D, E, F or G) per ml of solution, morepreferably from 100 to 2,500 LD50 units of botulinum neurotoxin complex(type A, B, C, D, E, F or G) or high purity botulinum neurotoxin (typeA, B, C, D, E, F or G) per ml of solution and most preferably from 100to 2,000 LD50 units of botulinum neurotoxin complex (type A, B, C, D, E,F or G) or high purity botulinum neurotoxin (type A, B, C, D, E, F or G)per ml of solution;

the concentration of surfactant will be from above critical micellarconcentration to a concentration of 1% v/v, and notably from about0.005% to 0.02% v/v in the case of polysorbate 80;

the concentration of crystalline agent will be from 0.1 to 0.5 M, morepreferably from 0.1 to 0.4 M, notably about 0.15 to 0.3 M; and

the concentration of buffer will be from 1 to 50 mM, more preferablyfrom 5 to 20 mM, notably about 10 mM.

As mentioned earlier, the solid or liquid pharmaceutical formulationaccording to the invention may contain a disaccharide. In that case, theconcentration of disaccharide in the solution to be lyophilised/theliquid pharmaceutical composition will be for example from 5 to 50 mM,preferably from 5 to 25 mM, more preferably from 10 to 20 mM, andnotably about 11.7 mM.

According to a preferred execution mode of the invention, the mixture ofthe different components of the pharmaceutical composition (i.e.botulinum neurotoxin complex (type A, B, C, D, E, F or G) or high puritybotulinum neurotoxin (type A, B, C, D, B, F or G), the surfactant andthe optional excipients such as the crystalline agent, the buffer or thedisaccharide) is lyophilised. The solid compositions thus obtained,which are also part of this invention, should preferably be stable forat least 12 months, more preferably for at least 18 months and in a moreparticularly preferred manner for at least 24 or even 36 months.

A composition according to the invention is considered stable during acertain period of time if at least 70% of the initial toxicity, asevaluated by assessing the LD₅₀ in mice or by any method validated withrespect to the LD₅₀ mouse assay (i.e. a method allowing a conversion ofits results into LD₅₀ units), is maintained over said period of time(cf. the part entitled “mouse toxicity assay” concerning the LD₅₀ mouseassay). Pharmaceutical compositions according to the invention can beused for preparing medicaments intended to treat a disease/a condition/asyndrome chosen from the following:

-   -   ophtalmological disorders selected from the group consisting of        blepharospasm, strabismus (including restrictive or myogenic        strabismus), amblyopia, oscillopsia, protective ptosis,        therapeutic ptosis for corneal protection, nystagmus, estropia,        diplopia, entropion, eyelid retraction, orbital myopathy,        heterophoria, concomitant misalignment, nonconcomitant        misalignment, primary or secondary esotropia or exotropia,        internuclear ophthalmoplegia, skew deviation, Duane's syndrome        and upper eyelid retraction;    -   movement disorders including hemifacial spasm, torticollis,        spasticity of the child or of the adult (e.g. in cerebral palsy,        post-stroke, multiple sclerosis, traumatic brain injury or        spinal cord injury patients), idiopathic focal dystonias, muscle        stiffness, Writer's cramp, hand dystonia, VI nerve palsy,        oromandibular dystonia, head tremor, tardive dyskinesia, tardive        dystonia, occupational cramps (including musicians' cramp),        facial nerve palsy, jaw closing spasm, facial spasm, synkinesia,        tremor, primary writing tremor, myoclonus,        stiff-person-syndrome, foot dystonia, facial paralysis,        painful-arm-and-moving-fingers-syndrome, tic disorders, dystonic        tics, Tourette's syndrome, neuromyotonia, trembling chin,        lateral rectus palsy, dystonic foot inversion, jaw dystonia,        Rabbit syndrome, cerebellar tremor, III nerve palsy, palatal        myoclonus, akasthesia, muscle cramps, IV nerve palsy,        freezing-of-gait, extensor truncal dystonia, post-facial nerve        palsy synkinesis, secondary dystonia, Parkinson's disease,        Huntington's chorea, epilepsy, off period dystonia, cephalic        tetanus, myokymia and benign cramp-fasciculation syndrome;    -   otorhinolaryngological disorders including spasmodic dysphonia,        hypersalivation, sialorrhoea, otic disorders, hearing        impairment, ear click, tinnitus, vertigo, Meniere's disease,        cochlear nerve dysfunction, stuttering, cricopharyngeal        dysphagia, bruxism, closure of larynx in chronic aspiration,        vocal fold granuloma, ventricular dystonia, ventricular        dysphonia, mutational dysphonia, trismus, snoring, voice tremor,        aspiration, tongue protrusion dystonia, palatal tremor, deep        bite of lip and laryngeal dystonia;    -   gastrointestinal disorders including achalasia, anal fissure,        constipation, temperomandibular joint dysfunction, sphincter of        Oddi dysfunction, sustained sphincter of Oddi hypertension,        intestinal muscle disorders, puborectalis syndrome, anismus,        pyloric spasm, gall bladder dysfunction, gastrointestinal or        oesophageal motility dysfunction, diffuse oesophageal spasm and        gastroparesis;    -   urogenital disorders including detrusor sphincter dyssynergia,        detrusor hyperreflexia, neurogenic bladder dysfunction (e.g. in        Parkinson's disease, spinal cord injury, stroke or multiple        sclerosis patients), bladder spasms, urinary incontinence,        urinary retention, hypertrophied bladder neck, voiding        dysfunction, interstitial cystitis, vaginismus, endometriosis,        pelvic pain, prostate gland enlargement (Benign Prostatic        Hyperplasia), prostatodynia, prostate cancer and priapism;    -   dermatological disorders including hyperhidrosis (including        axillary hyperhidrosis, palmar hyperhidrosis and Frey's        syndrome), bromhidrosis, cutaneous cell proliferative disorders        (including psoriasis), skin wounds and acne;    -   pain disorders including back pain (upper back pain, lower back        pain), myofascial pain, tension headache, fibromyalgia, painful        syndromes, myalgia, migraine, whiplash, joint pain,        post-operative pain, pain not associated with a muscle spasm and        pain associated with smooth muscle disorders;    -   inflammatory disorders including pancreatitis, neurogenic        inflammatory disorders (including gout, tendonitis, bursitis,        dermatomyositis and ankylosing spondylitis);    -   secretory disorders such as excessive gland secretions, mucus        hypersecretion and hyperlacrimation, holocrine gland        dysfunction;    -   respiratory disorders including rhinitis (including allergic        rhinitis), COPD, asthma and tuberculosis;    -   hypertrophic disorders including muscle enlargement, masseteric        hypertrophy, acromegaly and neurogenic tibialis anterior        hypertrophy with myalgia;    -   articular disorders including tennis elbow (or epicondilytis of        the elbow), inflammation of joints, coxarthrosis, hip        osteoarthritis, rotator muscle cap pathology of the shoulder,        rheumatoid arthritis and carpal tunnel syndrome;    -   endocrine disorders like type 2 diabetes, hyperglucagonism,        hyperinsulinism, hypoinsulinism, hypercalcemia, hypocalcemia,        thyroid disorders (including Grave's disease, thyroiditis,        Hashimoto's thyroiditis, hyperthyroidism and hypothyroidism),        parathyroid disorders (including hyperparathyroidism and        hypoparathyroidism), Cushing's syndrome and obesity;    -   autoimmune diseases like systemic lupus erythemotosus;    -   proliferative diseases including paraganglioma tumors, prostate        cancer and bone tumors;    -   traumatic injuries including sports injuries, muscle injuries,        tendon wounds and bone fractures; and    -   veterinary uses (e.g. immobilisation of mammals, equine colic,        animal achalasia or animal muscle spasms)

Pharmaceutical compositions according to the invention can also be usedfor cosmetic treatments including cosmetic treatments of the followingcosmetic disorders:

-   -   skin defects;    -   facial asymmetry;    -   wrinkles including glabellar frown lines and facial wrinkles;    -   downturned mouth;    -   hair loss; and    -   body odours.

Preferably, pharmaceutical compositions according to the invention willbe used for preparing medicaments intended to treat a disease/acondition/a syndrome chosen from the following:

-   -   ophtalmological disorders selected from the group consisting of        blepharospasm, strabismus (including restrictive or myogenic        strabismus), amblyopia, protective ptosis, therapeutic ptosis        for corneal protection and upper eyelid retraction;    -   movement disorders selected from the group consisting of        hemifacial spasm, torticollis, cerebral palsy spasticity of the        child, spasticity of the adult in post-stroke, multiple        sclerosis, traumatic brain injury or spinal cord injury        patients, idiopathic focal dystonias, muscle stiffness, Writer's        cramp, hand dystonia, VI nerve palsy, oromandibular dystonia,        head tremor, tardive dyskinesia, tardive dystonia, occupational        cramps (including musicians' cramp), facial nerve palsy, jaw        closing spasm, facial spasm, synkinesia, tremor, primary writing        tremor, myoclonus, stiff-person-syndrome, foot dystonia, facial        paralysis, painful-arm-and-moving-fingers-syndrome, tic        disorders, dystonic tics, Tourette's syndrome, neuromyotonia,        trembling chin, lateral rectus palsy, dystonic foot inversion,        jaw dystonia, Rabbit syndrome, cerebellar tremor, III nerve        palsy, palatal myoclonus, akasthesia, muscle cramps, IV nerve        palsy, freezing-of-gait, extensor truncal dystonia, post-facial        nerve palsy synkinesis, secondary dystonia, off period dystonia,        cephalic tetanus, myokymia and benign cramp-fasciculation        syndrome;    -   otorhinolaryngological disorders selected from the group        consisting of spasmodic dysphonia, hypersalivation, sialorrhoea,        ear click, tinnitus, vertigo, Meniere's disease, cochlear nerve        dysfunction, stuttering, cricopharyngeal dysphagia, bruxism,        closure of larynx in chronic aspiration, vocal fold granuloma,        ventricular dystonia, ventricular dysphonia, mutational        dysphonia, trismus, snoring, voice tremor, aspiration, tongue        protrusion dystonia, palatal tremor and laryngeal dystonia;    -   gastrointestinal disorders selected from the group consisting of        achalasia, anal fissure, constipation, temperomandibular joint        dysfunction, sphincter of Oddi dysfunction, sustained sphincter        of Oddi hypertension, intestinal muscle disorders, puborectalis        syndrome, anismus, pyloric spasm, gall bladder dysfunction,        gastrointestinal or oesophageal motility dysfunction, diffuse        oesophageal spasm, oesophageal diverticulosis and gastroparesis;    -   urogenital disorders selected from the group consisting of        detrusor sphincter dyssynergia, detrusor hyperreflexia,        neurogenic bladder dysfunction in Parkinson's disease, spinal        cord injury, stroke or multiple sclerosis patients, bladder        spasms, urinary incontinence, urinary retention, hypertrophied        bladder neck, voiding dysfunction, interstitial cystitis,        vaginismus, endometriosis, pelvic pain, prostate gland        enlargement (Benign Prostatic Hyperplasia), prostatodynia,        prostate cancer and priapism;    -   dermatological disorders selected from the group consisting of        axillary hyperhidrosis, palmar hyperhidrosis, Frey's syndrome,        bromhidrosis, psoriasis, skin wounds and acne;    -   pain disorders selected from the group consisting of upper back        pain, lower back pain, myofascial pain, tension headache,        fibromyalgia, myalgia, migraine, whiplash, joint pain,        post-operative pain and pain associated with smooth muscle        disorders;    -   inflammatory disorders selected from the group consisting of        pancreatitis, gout, tendonitis, bursitis, dermatomyositis and        ankylosing spondylitis;    -   secretory disorders selected from the group consisting of        excessive gland secretions, mucus hypersecretion and        hyperlacrimation and holocrine gland dysfunction;    -   respiratory disorders selected from the group consisting of        non-allergic rhinitis, allergic rhinitis, COPD and asthma;    -   hypertrophic disorders selected from the group consisting of        muscle enlargement, masseteric hypertrophy, acromegaly and        neurogenic tibialis anterior hypertrophy with myalgia;    -   articular disorders selected from the group consisting of tennis        elbow (or epicondilytis of the elbow), inflammation of joints,        coxarthrosis, hip osteoarthritis, rotator muscle cap pathology        of the shoulder, rheumatoid arthritis and carpal tunnel        syndrome;    -   endocrine disorders selected from the group consisting of type 2        diabetes, hypercalcemia, hypocalcemia, thyroid disorders,        Cushing's syndrome and obesity;    -   prostate cancer; and    -   traumatic injuries selected from the group consisting of sports        injuries, muscle injuries, tendon wounds and bone fractures;

or for performing cosmetic treatments wherein the cosmetic disorder tobe treated is selected from the group consisting of:

-   -   skin defects;    -   facial asymmetry;    -   wrinkles selected from glabellar frown lines and facial        wrinkles;

downturned mouth; and

-   -   hair loss.

More preferably, pharmaceutical compositions according to the inventionwill be used for preparing medicaments intended to treat a disease/acondition/a syndrome chosen from the following:

-   -   ophtalmological disorders selected from the group consisting of        blepharospasm and strabismus;    -   movement disorders selected from the group consisting of        hemifacial spasm, torticollis, cerebral palsy spasticity of the        child and arm or leg spasticity of the adult in post-stroke,        multiple sclerosis, traumatic brain injury or spinal cord injury        patients;    -   otorhinolaryngological disorders selected from the group        consisting of spasmodic dysphonia, hypersalivation, sialorrhoea,        cricopharyngeal dysphagia, bruxism, closure of larynx in chronic        aspiration, ventricular dystonia, ventricular dysphonia,        mutational dysphonia, trismus, snoring, voice tremor, tongue        protrusion dystonia, palatal tremor and laryngeal dystonia;    -   gastrointestinal disorders selected from the group consisting of        achalasia, anal fissure, constipation, temperomandibular joint        dysfunction, sphincter of Oddi dysfunction, sustained sphincter        of Oddi hypertension, intestinal muscle disorders, anismus,        pyloric spasm, gall bladder dysfunction, gastrointestinal or        oesophageal motility dysfunction and gastroparesis;    -   urogenital disorders selected from the group consisting of        detrusor sphincter dyssynergia, detrusor hyperreflexia,        neurogenic bladder dysfunction in Parkinson's disease, spinal        cord injury, stroke or multiple sclerosis patients, bladder        spasms, urinary incontinence, urinary retention, hypertrophied        bladder neck, voiding dysfunction, interstitial cystitis,        vaginismus, endometriosis, pelvic pain, prostate gland        enlargement (Benign Prostatic Hyperplasia), prostatodynia,        prostate cancer and priapism;    -   dermatological disorders selected from the group consisting of        axillary hyperhidrosis, palmar hyperhidrosis, Frey's syndrome,        bromhidrosis, psoriasis, skin wounds and acne;    -   pain disorders selected from the group consisting of upper back        pain, lower back pain, myofascial pain, tension headache,        fibromyalgia, myalgia, migraine, whiplash, joint pain,        post-operative pain and pain associated with smooth muscle        disorders;    -   inflammatory disorders selected from the group consisting of        pancreatitis and gout;    -   hyperlacrimation;    -   respiratory disorders selected from the group consisting of        non-allergic rhinitis, allergic rhinitis, COPD and asthma;

masseteric hypertrophy;

-   -   articular disorders selected from the group consisting of tennis        elbow (or epicondilytis of the elbow), inflammation of joints,        coxarthrosis, hip osteoarthritis, rotator muscle cap pathology        of the shoulder, rheumatoid arthritis and carpal tunnel        syndrome;    -   obesity;    -   traumatic injuries selected from the group consisting of muscle        injuries, tendon wounds and bone fractures;

or for performing cosmetic treatments wherein the cosmetic disorder tobe treated is selected from the group consisting of:

-   -   skin defects;    -   facial asymmetry;    -   wrinkles selected from glabellar frown lines and facial        wrinkles;    -   downturned mouth; and    -   hair loss.

In a particularly preferred manner, pharmaceutical compositionsaccording to the invention will be used for preparing medicamentsintended to treat a disease/a condition/a syndrome chosen from thefollowing: blepharospasm, hemifacial spasm, torticollis, cerebral palsyspasticity of the child and arm or leg spasticity of the adult inpost-stroke, multiple sclerosis, traumatic brain injury or spinal cordinjury patients, axillary hyperhidrosis, palmar hyperhidrosis, Frey'ssyndrome, skin wounds, acne, upper back pain, lower back pain,myofascial pain, migraine, tension headache, joint pain, tennis elbow(or epicondilytis of the elbow), inflammation of joints, coxarthrosis,hip osteoarthritis, rotator muscle cap pathology of the shoulder, muscleinjuries, tendon wounds and bone fractures;

or for performing cosmetic treatments wherein the cosmetic disorder tobe treated is selected from the group consisting of:

-   -   skin defects;    -   facial asymmetry; and    -   wrinkles selected from glabellar frown lines and facial        wrinkles.

The dose of botulinum neurotoxin complex (type A, B, C, D, E, F or G) orhigh purity botulinum neurotoxin (type A, B, C, D, E, F or G) whichshall be needed for the treatment of the diseases/disorders mentionedabove varies depending on the disease/disorder to be treated,administration mode, age and body weight of the patient to be treatedand health state of the latter, and it is the treating physician orveterinary that will eventually make the decision. Such a quantitydetermined by the treating physician or veterinary is called here“therapeutically efficient quantity”.

For botulinum neurotoxin complex (type A, B, C, D, E, F or G) or highpurity botulinum neurotoxin (type A, B, C, D, E, F or G), thistherapeutically efficient dose is often expressed as a function of thecorresponding LD₅₀. By LD₅₀ should be understood in the presentapplication the median intraperitoneal dose in mice injected withbotulinum neurotoxin complex (type A, B, C, D, E, F or G) or high puritybotulinum neurotoxin (type A, B, C, D, E, F or G) that causes death ofhalf of said mice within 96 hours.

The term “about” refers to an interval around the considered value. Asused in this patent application, “about X” means an interval from Xminus 10% of X to X plus 10% of X, and preferably an interval from Xminus 5% of X to X plus 5% of X.

Unless they are defined differently, all the technical and scientificterms used here have the same meaning as that usually understood by anordinary specialist in the field to which this invention belongs.Similarly, all publications, patent applications, all patents and allother references mentioned here are incorporated by way of reference.

The following examples are presented by way of illustration and shouldin no way be considered to limit the scope of the invention.

EXAMPLES Example 1 A Liquid Pharmaceutical Composition Containing theFollowing Components is Prepared

Clostridium botulinum type A1 neurotoxin complex 2,000 LD₅₀ units/mlSucrose 11.7 mM Histidine 10 mM Sodium chloride 0.3M Polysorbate 800.01% v/v pH 6.5

The mixture containing nominally 2,000 LD₅₀ units of botulinum toxin perml is lyophilised in a sterilised vial, which is then sealed. The solidcomposition obtained is stable for at least 18 months when stored at atemperature between 2 and 8° C. and at least 6 months at 23 to 27° C.

Example 2 A Liquid Pharmaceutical Composition Containing the FollowingComponents is Prepared

Clostridium botulinum type A1 neurotoxin complex 500 LD₅₀ units/mlSucrose 11.7 mM Histidine 10 mM Sodium chloride 0.3M Polysorbate 800.01% v/v PH 6.5

The liquid composition thus prepared is sealed in a syringe type devicewith no liquid/gaseous interface. Stored in these conditions, it isstable for at least six months at 23 to 27° C. and at least twelvemonths at 2-8° C.

Example 3 A Liquid Pharmaceutical Composition Containing the FollowingComponents is Prepared

Clostridium botulinum type A1 neurotoxin complex 500 LD₅₀ units/mlSucrose 11.7 mM Histidine 10 mM Sodium chloride 0.15M Polysorbate 800.01% v/v PH 6.5

The liquid composition thus prepared is sealed in a syringe type devicewith no liquid/gaseous interface. Stored in these conditions, it isstable for at least six months at 23 to 27° C. and at least twelvemonths at 2-8° C.

Analytical Methods Mouse Toxicity Assay

A mouse toxicity assay can be used to measure the toxicity of botulinumneurotoxin complex (type A, B, C, D, E, F or G) or high purity botulinumneurotoxin (type A, B, C, D, E, F or G). In the assay, a standarddiluent will be used to prepare a range of dilutions at or about theestimated LD₅₀ value. The range and scale of dilutions is arranged so asto establish an accurate LD₅₀ value.

Mice are injected intraperitoneally with a known and standardised volumeof diluted toxin. After 96 hours, the number of deaths and survivors ineach dilution group will be recorded. The LD₅₀ value is the median dosewhich kills half of the injected animals within 96 hours.

A composition according to the invention is considered stable over acertain period of time if at least 70% of the initial toxicity ismaintained over said period of time relative to a reference preparation.

1. A solid or liquid pharmaceutical composition comprising: (a)botulinum neurotoxin complex (type A, B, C, D, E, F or G) or high puritybotulinum neurotoxin (type A, B, C, D, E, F or G), and (b) a surfactant,2. A solid or liquid pharmaceutical composition according to claim 1further comprising a crystalline agent.
 3. A solid or liquidpharmaceutical composition according to claim 2 wherein the crystallineagent is sodium chloride.
 4. A solid or liquid pharmaceuticalcomposition according to one of claims 1 to 3 further comprising abuffer to maintain pH between 5.5 to 7.5.
 5. A solid or liquidpharmaceutical composition according claim 4 wherein the buffer ismaintaining a pH from 5.8 to 7.0.
 6. A solid or liquid pharmaceuticalcomposition according to any of claims 1 to 5 further comprising adisaccharide.
 7. A solid or liquid pharmaceutical composition accordingto claim 6 wherein the disaccharide is chosen from the group consistingof sucrose, trehalose, lactose and mannitol.
 8. A solid or liquidpharmaceutical composition according to any of claims 1 to 7 whichcontains botulinum neurotoxin complex type A.
 9. A pharmaceuticalcomposition according to any of claims 1 to 7 which contains high puritybotulinum neurotoxin type A.
 10. A pharmaceutical composition accordingto any of claims 1 to 9 in which the surfactant is polysorbate
 80. 11. Asolid or liquid pharmaceutical composition substantially as hereindescribed.